How to calibrate pgFocus
- PgFocus must be connected to a piezo controller, which is connected to a piezo positioner.
- PgFocus assumes that 1 Volt moves the objective 10µm.
- Focus on a coverslip as you would normally.
- Cells are not needed but the media should be same.
- A focus position must be able to be detected by pgFocus, and it should be roughly centered on the Linear Light Array (LLA). Anywhere between 50-70 offset should be fine.
- In the Stand-alone pgFocus application, type the letter 'c' to activate the calibration process.
- You can also use the pgFocus µManager plugin.
- PgFocus will move the objective until the focus position reaches 100 offset on the LLA.
- This will determine the maximum voltage that pgFocus will use during the calibration process.
- pgFocus will either use the maximum voltage as determined by this step, or voltage needed to move the objective 3000nm up, whichever is less.
- PgFocus will then slowly lower the objective by 500nm until it reaches -3000 nm or the bottom the linear array sensor (28 offset).
- Voltage and LLA offset will be collected at each step.
- PgFocus will then perform a linear regression on the data points it collected, and save the result in the eprom.
500 nm is hard coded based on DAUPERVOLT/MICRONPERVOLT/2 = 81.9. It shouldn't be hardcoded... : D